WO2023159532A1 - Procédé de détection de pesticides organophosphorés à l'aide d'une puce microfluidique basée sur un film de détection fluorescent - Google Patents

Procédé de détection de pesticides organophosphorés à l'aide d'une puce microfluidique basée sur un film de détection fluorescent Download PDF

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WO2023159532A1
WO2023159532A1 PCT/CN2022/078143 CN2022078143W WO2023159532A1 WO 2023159532 A1 WO2023159532 A1 WO 2023159532A1 CN 2022078143 W CN2022078143 W CN 2022078143W WO 2023159532 A1 WO2023159532 A1 WO 2023159532A1
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channel
detection
solution
fluorescent sensing
microfluidic
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Chinese (zh)
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胡雪桃
邹小波
蒋培
石吉勇
张新爱
黄晓玮
李志华
李亚惠
李文亭
张俊俊
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江苏大学
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    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G01N21/643—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" non-biological material
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502707—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the manufacture of the container or its components
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y15/00—Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y30/00—Nanotechnology for materials or surface science, e.g. nanocomposites
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y40/00—Manufacture or treatment of nanostructures
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0627—Sensor or part of a sensor is integrated
    • B01L2300/0654—Lenses; Optical fibres
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N2021/6417—Spectrofluorimetric devices
    • G01N2021/6421—Measuring at two or more wavelengths
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the invention belongs to the field of detection of organophosphorus pesticides, in particular to a method for detecting organophosphorus pesticides with a microfluidic chip based on a fluorescent sensing film.
  • Organophosphorus pesticides refer to organic compounds containing phosphorus elements, such as common organophosphorus pesticides trichlorfon, parathion, dimethoate and dichlorvos, etc., which have the effect of preventing and controlling crop diseases, insect pests and weeds.
  • the above methods can achieve accurate detection of organophosphorus pesticides, but there are still disadvantages such as cumbersome operation or dependence on large and heavy instruments, and cannot realize rapid and portable detection of pesticide residues on site.
  • fluorescent nanomaterials show great application potential in the fields of labeling, tracing, imaging and detection due to their stable morphological structure and high luminous efficiency.
  • detection of pesticide residues based on fluorescent nanomaterials is devoted to the preparation of specific nanomaterials, and the detection must be carried out in solution.
  • the commonly used specific molecules for detection are biological molecules such as aptamers and antigens/antibodies.
  • Fluorescent nanomaterials In the solution, due to poor solubility or poor stability, nanoparticle precipitation and fluorescence quenching will occur. The application of biomolecules leads to high detection cost and poor detection repeatability. The disadvantages of unstable solution and difficult portability limit the use of fluorescent nanomaterials. Application in field testing.
  • the combination of other materials or other detection methods can play a greater detection role, such as the preparation of fluorescent sensing films and the combination of microfluidic chips.
  • the disadvantages of poor stability in solution and easy photobleaching of fluorescent nanomaterials can be overcome by preparing fluorescent thin film sensors, and the sensors are endowed with characteristics such as easy portability and easy use.
  • the microfluidic chip technology is used to integrate various processes involved in sample pretreatment, mixing reaction and fluorescence detection into a chip of several square centimeters, which has multi-functional integration, small size and It is easy to carry and other advantages, and it is a commonly used instant detection device.
  • the present invention constructs a H 2 O 2 fluorescence
  • the sensing film provides an organophosphorus pesticide microfluidic chip system and detection method that integrates automatic sampling, pretreatment and detection functions, and intends to solve the problem that the microfluidic chip cannot be standardized and standardized to detect organophosphorus pesticides, and The problem of being unable to realize rapid on-site detection of organophosphorus pesticides.
  • a microfluidic chip system based on a fluorescent sensing film and an organophosphorus pesticide detection method constructed by the present invention are characterized in that they include the preparation of a specific fluorescent sensing film, the construction of a microfluidic chip system, the establishment of an on-site detection method, and the actual application method.
  • Step 1 Construction of porous fluorescent sensing film, including the synthesis of specific fluorescent materials and the construction of fluorescent sensing film, which can be used for the specific detection of acetylcholine hydrolyzate H2O2 :
  • molybdenum disulfide nanosheets, polystyrene polymers, and polydimethyldiallylammonium chloride were dissolved in ethanol at a certain ratio to obtain a mixed solution, and then the fluorescent sensing layer modified quartz substrate was mixed Soak in the liquid for M times, form M layers of macromolecular barrier layer on the surface of the fluorescent sensing layer, and finally obtain the organophosphorus pesticide fluorescent sensing film comprising the fluorescent sensing layer and the macromolecular barrier layer, which is recorded as a porous fluorescent sensing film;
  • Step 2 Design a microfluidic chip with automatic sample injection, sample pretreatment, sample and reagent mixing reaction, and detection functions;
  • the microfluidic chip includes a sample channel, a reaction tank, a detection tank, a microfluidic channel, and an optical fiber channel; there are n reaction tanks, which are sequentially recorded as the first reaction tank, the second reaction tank..., the n-1th reaction tank and the nth reaction tank;
  • One end of the microfluidic chip has a sample port, and one end of the first reaction tank is provided with a sample channel connected to the sample port; the other end is connected to the second reaction tank..., the n-1 reaction tank and the nth reaction tank through the microfluidic channel
  • the reaction tanks are connected sequentially; the other end of the nth reaction tank is connected to one end of the detection tank through a microfluidic channel; the other end of the detection tank is provided with a fiber optic channel, and is not connected to the fiber optic channel;
  • sampling channels on one side of the microfluidic chip, which are sequentially recorded as the first sampling channel, the second sampling channel..., the n-1th sampling channel and the nth sampling channel; the first The sampling channel communicates with the first reaction tank, the second sampling channel communicates with the second reaction tank..., the n-1th sampling channel communicates with the n-1th reaction tank; the nth sampling channel Connected with the nth reaction tank;
  • a fluorescent sensing thin film pick-and-release port and a visual detection port are opened above the detection groove; the n is a positive integer.
  • the preparation method of the microfluidic chip is:
  • the channel structure of the microfluidic chip First design the channel structure of the microfluidic chip, and print out the microfluidic channel model with the dissolving support material as the base material; then transfer the microfluidic channel model to the container, pour polydimethylsiloxane (PDMS) and The complex formed by mixing the curing agent is heated to a certain temperature to harden the PDMS to obtain a microfluidic platform template, and the microfluidic channel model in the microfluidic platform template is dissolved by an aqueous solution of an organic solvent to obtain a microfluidic chip platform; for microfluidic chips The upper surface of the platform is pretreated so that the microfluidic channel on the upper surface communicates with the external environment;
  • PDMS polydimethylsiloxane
  • Step 3 the establishment of organophosphorus pesticide fluorescence color card and quantitative detection model, the specific steps are:
  • microfluidic chip system for organophosphorus pesticides mainly including constant pressure injection pumps, syringes, connecting tubes, microfluidic chips, fluorescent optical fibers, lasers, spectrometers, signal transmitters, and signal displays;
  • the constant-pressure syringe pump is connected to the sampling port of the microfluidic chip through a connecting tube for sampling;
  • the syringe is connected to the sampling channel through the connecting tube, and the reaction reagent is added into the reaction tank; the reaction reagent is used to degrade acetylcholine to generate H 2 O 2 ;
  • One end of the fluorescent optical fiber is placed in the optical fiber channel to obtain the fluorescent signal of the porous fluorescent sensing film in the detection groove, and the other end is connected to the laser and the spectrometer; the spectrometer transmits the obtained porous fluorescent sensing film signal through the signal transmitter transmission to the signal display;
  • the amount ratio of europium nitrate, pyromellitic acid, oxalic acid and acetonitrile-ethanol solution in S1 of step 1 is 0.1 ⁇ 10mmol:0.05 ⁇ 0.5mmol:0.05 ⁇ 0.5mmol:50 ⁇ 100mL; acetonitrile-ethanol solution It is a mixture of acetonitrile and ethanol, wherein the mixed volume ratio of acetonitrile and ethanol is 6-10:1-4; the reaction temperature under the certain temperature conditions is 100-200°C, and the reaction time is 12-72h;
  • the purification step is to use an organic solvent to repeatedly wash 3 to 10 times, and the organic solvent includes any one of N,N-dimethylformamide, ethanol or acetone; the drying refers to drying at 40 to 60°C until constant weight;
  • the optimum excitation wavelengths of the organometallic framework powder and the organometallic framework material solution are consistent, ranging from 500 to 650 nm, and the optimum emission wavelengths are W 1 , W 2
  • the mass concentration of the oxalate solution in S1 of step 1 is 0.5-1 mg/mL; the mass concentration of the organometallic framework solution is 2-10 mg/mL; the oxalate solution and the organometallic framework
  • the volume ratio of the solution is 1:1; the oxalate solution is mixed with the organometallic framework material solution, and the temperature of oscillation under certain temperature conditions is 30-50°C, and the oscillation time is 12-48h; the oxalate-
  • the concentration of the organic metal framework material is 2-10 mg/mL, the concentration of the potassium chloroplatinite is 0.3-5 mg/mL, the concentration of the sodium borohydride is 0.1-1 mg/mL; the oxalate-organic
  • the volume ratio of metal frame material, potassium chloroplatinite and sodium borohydride is 1:0.1 ⁇ 2:0.5 ⁇ 1.5;
  • the particle size of the platinum nanoparticles in the Pt@TCPO-EuMOF solution does not exceed 50nm.
  • the cleaning of the quartz substrate in S2 of step 1 is specifically ultrasonic cleaning in an ethanol solution for 3 to 8 times, each cleaning for 1 to 10 minutes; the polydimethyldiallylammonium chloride (PDADMA) solution
  • PDADMA polydimethyldiallylammonium chloride
  • concentration is 0.5-5g/L
  • concentration of Pt@TCPO-EuMOF solution is 0.5-10mg/mL.
  • the soaking time in both PDADMA solution and Pt@TCPO-EuMOF solution is 5-30min.
  • the number of times N of repeated immersion is 5 ⁇ 100;
  • the consumption ratio of described molybdenum disulfide nanosheet, polystyrene polymer, polydimethyldiallylammonium chloride and ethanol is 0.1 ⁇ 5mg:0.2 ⁇ 2mg: 0.01-0.1mg: 1ml; the soaking times M is 2-10, and the soaking time is 3-10min; the macromolecule barrier layer on the surface of the porous fluorescent sensing film has a pore size of 2-15nm.
  • the channel structure of the microfluidic chip is designed as described in step 2.
  • the channel structure includes a microfluidic channel, a sample channel, a reaction tank, and a detection tank.
  • the specific design requirements are: the total length of the microfluidic channel is 5 to 20 cm, and the channel The depth and width are both 0.4-4mm; the lengths of the sample channel, the first sampling channel, the second sampling channel..., the n-1th sampling channel and the nth sampling channel are all 0.8-2cm, and the depth and The widths are all 0.4-5mm, the depths of the first reaction tank, the second reaction tank..., the n-1th reaction tank, the nth reaction tank and the detection tank are all 0.5-5mm; the area of the reaction tank is 0.5 ⁇ 3cm 2 ; the area of the detection groove is 0.3 ⁇ 5cm 2 ; the dimensions of the microfluidic chip and the quartz substrate are consistent, and the length, width and height are 6 ⁇ 15cm, 1 ⁇ 10cm and 0.5 ⁇ 2cm
  • the soluble support material described in step 2 is one of polyoxymethylene, polyvinyl alcohol or acrylic copolymer, or a mixture of multiple arbitrary proportions; the polydimethylsiloxane and its curing agent The mass ratio is 7-9:1-3; the curing agent is a type of silane coupling reagent, mainly composed of vinyltriamine, aminoethylpiperazine, diaminodiphenylmethane and m-phenylenediamine Or a mixture of several kinds; the organic solvent aqueous solution refers to a mixed solution of water and an organic solvent with a volume ratio of 1-4:6-9; the organic solvent includes one or more of acetone, isopropanol or nitric acid; The heating to a certain temperature is 65-80°C.
  • the aperture of the fluorescent sensing film access port and the visual detection port is 0.6-2.1 cm; the detection groove is not connected to the optical fiber channel and the distance is 0.5-1 cm, and the length, width, and height of the optical fiber channel are 1-5 cm respectively.
  • the reaction reagent in step three includes one or more of acetylcholinesterase, acetylcholine or choline oxidase; the fluorescent optical fiber is specifically a 1*2 type fluorescent optical fiber.
  • the reaction period in the detection tank described in step 3 is 2 to 10 minutes; the wavelength of the ultraviolet lamp and the excitation light is 301 to 399 nm; the concentration range of the standard organophosphorus pesticide is 0 to 1 g/mL, respectively record are 0, n 1 , n 2 , n 3 ,..., n h ; the pictures of the fluorescent sensing film are recorded as p 0 , p 1 , p 2 , p 3 ,..., ph respectively; the wavelength is W 1
  • the fluorescence signal intensities corresponding to different concentrations of organophosphorus pesticide standard substances are recorded as I 1,0 , I 1.1 , I 1.2 , I 1.3 , ..., I 1.h , and the wavelengths are determined by the different concentrations of organophosphorus pesticide standard substances at W 2
  • the corresponding fluorescent signal intensities are recorded as I 2.0 , I 2.1 , I 2.2 , I 2.3 , ..., I 2.h , ..., and the
  • the sum of the fluorescence signal intensity difference is y, which is respectively recorded as y 1 , y 2 , y 3 , ..., y h ;
  • the period of time for the reaction in the detection tank in step 4 is 2-10 minutes; the reaction reagent includes one or more of acetylcholinesterase, acetylcholine or choline oxidase.
  • the present invention constructs a fluorescent organometallic framework material with a cage structure, fixes specific molecular oxalate in the pore structure, and simultaneously introduces noble metal nanoparticles to form an oxalate-organometallic framework supported by platinum nanoparticles
  • Composite materials in which platinum nanoparticles can catalyze the specific reaction between oxalate and H 2 O 2 , and the chemiluminescence generated by the reaction can specifically enhance the fluorescence signal of the organometallic framework, and the oxalate-based
  • the new chemiluminescence resonance energy transfer system of H 2 O 2 reaction products and EuMOF the prepared fluorescent sensing material has dual functions of catalytic reaction and specific detection, which can improve the specificity and sensitivity of detecting organophosphorus pesticides.
  • the present invention assembles platinum nanoparticles@oxalate-organic metal framework composite material on the surface of the quartz substrate to prepare the fluorescent sensing layer.
  • Fluorescent materials are easy to use and portable.
  • the porous two-dimensional nanosheet material is wrapped on the outer surface of the fluorescent sensing layer, which can block macromolecules such as protease contained in the reaction solution from entering the fluorescent sensing layer, and reduce the impact of macromolecules such as protease on the fluorescent sensing layer. Unfavorable effects of fluorescent signal, improve detection accuracy and reduce detection limit.
  • the present invention builds a portable organophosphorus pesticide microfluidic chip system by designing a microfluidic chip with functions such as automatic sampling, sample pretreatment, and mixed reaction between samples and detection reagents, so that the detection equipment is integrated and miniaturized.
  • the standardization and standardization of pesticide testing will ultimately improve the stability and reproducibility of on-site testing.
  • the present invention combines the fluorescent nano-film with the microfluidic chip, and by designing the specific fluorescent film, the reaction between the organophosphorus pesticide and the specific detection reagent is sealed in the stable microfluidic chip, and the fluorescent signal can be obtained with the naked eye.
  • the change of organophosphorus pesticides can be quickly visualized, and the fluorescence intensity can be obtained by a portable spectrometer, so that the rapid quantitative detection of organophosphorus pesticides can be realized.
  • Figure 1 is a schematic diagram of the structure of the porous fluorescent sensing film, in which: 1-quartz substrate, 2-polydimethyldiallylammonium chloride layer, 3-platinum nanoparticles@oxalate-organic metal framework composite material layer, 4-molybdenum disulfide nanosheet-polystyrene polymer porous barrier layer;
  • Figure 2 is a schematic diagram of the microfluidic chip structure, wherein: 1-sample channel, 2-first reaction tank, 3-first microfluidic channel, 4-second reaction tank, 5-second microfluidic channel, 6-third Reaction tank, 7-third microfluidic channel, 8-detection tank, 9-fluorescent sensing film access port and visual detection port, 10-fiber optic channel, 11-quartz substrate, 12-first sampling channel, 13 -the second sampling channel, 14-the third sampling channel, 15-microfluidic platform;
  • Fig. 3 is a flow chart of organophosphorus pesticide detection; wherein 1-constant pressure injection pump, 2-first connecting pipe, 3-first syringe, 4-second syringe, 5-third syringe, 6-second connecting pipe, 7-third connecting tube, 8-fourth connecting tube, 9-microfluidic chip, 10-1*2 type fluorescent optical fiber, 11-laser, 12-spectrometer, 13-signal transmitter, 14-signal display.
  • a microfluidic chip preparation and on-site detection method for organophosphorus pesticides comprising the following steps:
  • the organic metal framework material (EuMOF) is obtained and placed in a 45°C drying oven Dry to obtain powder, redissolve in water to obtain organometallic framework solution, determine the optimal excitation wavelength of the two by obtaining the fluorescence spectrum of the solution and powder is 560nm, the excitation wavelength range is 500-650nm, and the optimal emission wavelength is 520nm and 610nm, Under the irradiation of the best excitation light (560nm), both the organometallic framework solution and the powder show weak red fluorescence, because the ligand has a large number of carboxyl groups, so the organometallic framework material is negatively charged; 10mL of bis( 2,4,6-Trichlorophenyl)oxalate solution (TCPO) was stirred with 10mL organometallic framework solution with a concentration of 5mg/mL for 48h to obtain bis(2,4,6-trichlorophenyl)oxalate -Organometallic framework composite solution; the
  • the microfluidic chip includes a sample channel 1, a first reaction tank 2, a first microfluidic channel 3, a second reaction tank 4, a second microfluidic channel 5, a third reaction tank 6, and a third microfluidic channel 7.
  • Groove 8 Fluorescent sensing film access port and visual detection port 9, Fiber channel 10, Quartz substrate 11, First sampling channel 12, Second sampling channel 13, Third sampling channel 14 and Microfluidic platform 15 ;
  • One end of the microfluidic chip opens a sample port and communicates with the first reaction tank 2 through the sample channel 1; the other end of the first reaction tank 2 communicates with the second reaction tank 4 through the first microfluidic channel 3; the second reaction tank The other end of 4 communicates with the third reaction tank 6 through the second microfluidic channel 5; the other end of the third reaction tank 6 communicates with the detection tank 8 through the third microfluidic channel 7; the right side of the detection tank Fiber Channel is provided and is not connected to Fiber Channel;
  • One side of the microfluidic chip is also provided with 3 sampling channels, which are successively recorded as the first sampling channel 12, the second sampling channel 13 and the third sampling channel 14;
  • a reaction tank 2 is communicated, the second sampling channel 13 is communicated with the second reaction tank 4, and the third sampling channel 14 is communicated with the third reaction tank 6;
  • the preparation method of the microfluidic chip is:
  • microfluidic design diagram comprising 1 sample channel, 3 injection channels, 3 circular reaction slots, 1 circular detection slot and 3 U-shaped microfluidic channels), 3 U-shaped microfluidic channels
  • the total length and width of the fluid channel are 9cm and 3cm respectively, the depth of the channel is 500 ⁇ m, the width of the microfluidic channel itself is 1mm, and the length and width of the sample channel and the sampling channel are 1cm and 0.5cm respectively;
  • Alcohol composite materials are used as raw materials, and microfluidic channel templates are printed by 3D printing technology;
  • quartz substrate 11 is a transparent material, through which the quartz The substrate 11 can clearly see the substance and its flow in the pipeline;
  • a fluorescent sensing film pick-and-place port and a visual detection port 9 are provided for the pick-and-place of the fluorescent sensing film and the acquisition of the fluorescent visualization signal; a strip is processed on the right side of the detection groove 8
  • the optical fiber channel 10 that is not connected with the detection groove 8 is 0.1 cm away from the detection groove 8, and finally obtains a microfluidic chip;
  • Sample channel 1 is the entry channel of the organophosphorus pesticide sample
  • the first sampling channel 12, the second sampling channel 13, and the third sampling channel 14 are respectively the sampling channels of acetylcholinesterase inlet, acetylcholine inlet and choline oxidase .
  • Step 3 establishment of microfluidic chip detection method
  • the present invention first builds a standard detection system, as shown in Figure 3, including a constant pressure injection pump 1, a first connecting pipe 2, a first syringe 3, a second syringe 4, a third syringe 5, a second connecting pipe 6,
  • the constant pressure syringe pump 1 is connected to the sampling port of the microfluidic chip 9 through the first connecting pipe 2;
  • the first syringe 3, the second syringe 4, and the third syringe 5 pass through the second connecting pipe 6, the third connecting pipe 7, the fourth connecting pipe 8 and the first sample injection channel 12 and the second sample injection channel 12 of the microfluidic chip 9 respectively.
  • Channel 13 is connected to the third sampling channel 14;
  • the 1*2 type fluorescent optical fiber 10 is used to obtain the fluorescent signal of the porous fluorescent sensing film in the detection groove 8, and the other end is connected with the laser 11 and the spectrometer 12;
  • the sensitive film signal is transmitted to the signal display 14;
  • the steps to establish a detection method for organophosphorus pesticides mainly include:

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Abstract

Procédé de détection de pesticides organophosphorés à l'aide d'une puce microfluidique (9) basée sur un film de détection fluorescent, appartenant au domaine de la détection de pesticides organophosphorés. Tout d'abord, un film de détection fluorescent poreux et une puce microfluidique (9) sont construits, le film de détection fluorescent étant préparé au moyen d'un auto-assemblage couche par couche d'une couche de matériau composite à structure organique de nanoparticules de platine à oxalate-métal (3) et de nanofeuilles bidimensionnelles poreuses, et ayant les fonctions de détection spécifique de pesticides organophosphorés et de blocage d'interférants macromoléculaires. La puce microfluidique (9) est constituée d'un canal d'échantillon (1), d'un canal d'entrée d'échantillon, de rainures de réaction, de canaux microfluidiques, d'une rainure de détection (8) et d'un canal de fibre optique (10), de telle sorte que des processus de prétraitement et de détection d'échantillon sont intégrés dans la puce ; et, en combinaison avec une pompe d'injection à pression constante portative (1), un laser (11), un spectrographe (12), un émetteur de signal (13) et une unité d'affichage de signal (14), un système de détection de pesticides organophosphorés est construit, de telle sorte que le dispositif de détection est miniaturisé et intégré, et la détection de pesticides organophosphorés est standardisée. Par accouplement du nano-film et de la puce microfluidique (9), une détection qualitative et quantitative sur site rapide de pesticides organophosphorés est obtenue.
PCT/CN2022/078143 2022-02-24 2022-02-28 Procédé de détection de pesticides organophosphorés à l'aide d'une puce microfluidique basée sur un film de détection fluorescent WO2023159532A1 (fr)

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CN115850980B (zh) * 2022-11-22 2024-04-05 厦门华厦学院 一种温度传感材料及其制备方法和应用
CN117244599B (zh) * 2023-11-15 2024-02-02 湘潭大学 一种pdms微流控芯片制作方法

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